Anti-VPS35 抗体 [EPR11501(B)] (ab157220)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR11501(B)] to VPS35
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Human
Related conjugates and formulations
製品の概要
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製品名
Anti-VPS35 antibody [EPR11501(B)]
VPS35 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR11501(B)] to VPS35 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
適用なし: IP -
種交差性
交差種: Mouse, Human
交差が予測される動物種: Rat -
免疫原
Synthetic peptide within Human VPS35. The exact sequence is proprietary.
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ポジティブ・コントロール
- Flow Cyt (intra): MOLT4 cells. WB: HeLa, HepG2, Raji, MOLT4, wild-type HAP1, NIH/3T3 and A549 cell lysates. IHC-P: Human kidney and human ovarian carcinoma tissue. ICC/IF: Wild-type HAP1 cells.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at -20°C. -
バッファー
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 9% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR11501(B) -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab157220の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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Flow Cyt (Intra) |
1/10 - 1/100.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/10000 - 1/50000. Predicted molecular weight: 92 kDa.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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ICC/IF |
Use a concentration of 0.2 µg/ml.
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特記事項 |
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Flow Cyt (Intra)
1/10 - 1/100. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000 - 1/50000. Predicted molecular weight: 92 kDa. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
Use a concentration of 0.2 µg/ml. |
ターゲット情報
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機能
Essential component of the retromer complex, a complex required to retrieve lysosomal enzyme receptors (IGF2R and M6PR) from endosomes to the trans-Golgi network. Also required to regulate transcytosis of the polymeric immunoglobulin receptor (pIgR-pIgA). -
組織特異性
Ubiquitous. Highly expressed in heart, brain, placenta, skeletal muscle, spleen, thymus, testis, ovary, small intestine, kidney and colon. -
配列類似性
Belongs to the VPS35 family. -
細胞内局在
Cytoplasm. Membrane. - Information by UniProt
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参照データベース
- Entrez Gene: 100133770 Human
- Entrez Gene: 55737 Human
- Entrez Gene: 65114 Mouse
- Entrez Gene: 25479 Rat
- Omim: 601501 Human
- SwissProt: Q96QK1 Human
- SwissProt: Q9EQH3 Mouse
- Unigene: 454528 Human
see all -
別名
- DKFZp434E1211 antibody
- DKFZp434P1672 antibody
- FLJ10752 antibody
see all
画像
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ab157220 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab157220 at 0.2 µg/mL and ab7291 at 1 µg/mL overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) (shown in pseudo colour green) and goat secondary antibody to Mouse IgG (Alexa Fluor® 594) (ab150120) (shown in pseudo colour red) both at 1/1000. Nuclear DNA was labelled with DAPI (shown in pseudo colour blue).
Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: VPS35 knockout HAP1 cell lysate (20 µg)
Lane 3: NIH/3T3 cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab157220 observed at 91 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab157220 was shown to specifically react with VPS35 in wild-type HAP1 cells. No band was observed when VPS35 knockout samples were examined. Wild-type and VPS35 knockout samples were subjected to SDS-PAGE. ab157220 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging. -
ab157220 staining VPS35 in wild-type HAP1 cells (top panel) and VPS35 knockout HAP1 cells (bottom panel). The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab157220 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudo colour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
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All lanes : Anti-VPS35 antibody [EPR11501(B)] (ab157220) at 1/10000 dilution
Lane 1 : HeLa cell lysate
Lane 2 : HepG2 cell lysate
Lane 3 : Raji cell lysate
Lane 4 : MOLT4 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-rabbit HRP at 1/2000 dilution
Predicted band size: 92 kDa -
Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling VPS35 with ab157220 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human ovarian carcinoma tissue labeling VPS35 with ab157220 at 1/50 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of permeabilized MOLT4 cells labeling VPS35 with ab157220 at 1/10 dilution (red) or a rabbit IgG (negative) (green).
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (22)
ab157220 は 22 報の論文で使用されています。
- Roşianu F et al. Loss of NDR1/2 kinases impairs endomembrane trafficking and autophagy leading to neurodegeneration. Life Sci Alliance 6:N/A (2023). PubMed: 36446521
- Tu H et al. Increased expression of pathological markers in Parkinson's disease dementia post-mortem brains compared to dementia with Lewy bodies. BMC Neurosci 23:3 (2022). PubMed: 34983390
- Courtellemont T et al. CROP: a retromer-PROPPIN complex mediating membrane fission in the endo-lysosomal system. EMBO J 41:e109646 (2022). PubMed: 35466426
- Niu M et al. VPS35 D620N knockin mice recapitulate cardinal features of Parkinson's disease. Aging Cell 20:e13347 (2021). PubMed: 33745227
- Liu Y et al. Depletion of VPS35 attenuates metastasis of hepatocellular carcinoma by restraining the Wnt/PCP signaling pathway. Genes Dis 8:232-240 (2021). PubMed: 33997170